Cyclohexene

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 3 to 9 November, 2010

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Cyclohexene

EC Number:

203-807-8

EC Name:

Cyclohexene

Cas Number:

110-83-8

Molecular formula:

C6H10

IUPAC Name:

cyclohexene

Details on test material:

Lot No. 10817K
Purity: 99.83%

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: ELEVAGE JANVIER
Route des Chènes Secs B.P. 4105 53940 LE GENEST-ST-ISLE, France
- Age at study initiation: 11-12 weeks old (age-matched, within one week)
- Weight at study initiation: 23.3 – 25.6 grams (The weight variation in animals involved in the study did not exceed ± 20 % of the mean weight)
- Housing: Group caging / mice were provided with glass tunnel-tubes

- Diet (e.g. ad libitum): Animals received ssniff SM R/M-Z+H "Autoclavable complete diet for rats and mice – breeding and maintenance" (Batch number: 847 4257, Expiry Date: January 2011) produced by ssniff Spezialdiäten GmbH (Ferdinand-Gabriel-Weg 16, D-59494 Soest, Germany), ad libitum. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

- Water (e.g. ad libitum): Animals received tap water from the municipal supply from 500 mL bottle, ad libitum. Water quality control analysis was performed once every three months and microbiological assessment was performed monthly by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József Attila u. 36., Hungary).

- Acclimation period: 26 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 15-20 air exchange/hour
- Photoperiod (hrs light): 12 hours daily

IN-LIFE DATES: From: To: from 6.00 a.m. to 6.00 p.m.

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

Test item concentration are 100, 50, 25 (% w/v), respectively.

No. of animals per dose:

4 animals / treatment group

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: 100% (undiluted) and 50 (w/v) % in AOO
- Irritation: No irritation was observed. Doses of 100 (undiluted) and 50 (w/v) % were considered to be acceptable for the main tests.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: The stimulation index values were 2.7, 1.2 and 1.4 at concentrations of 100 %, 50 % and 25 %, respectively.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: The DPM values were 1748.5, 823.5 and 949.5 at concentrations of 100 %, 50 % and 25 %, respectively.

Any other information on results incl. tables

- CLINICAL OBSERVATION

No mortality or signs of systemic toxicity were observed during the study.

- BODY WEIGHT MEASUREMENT

No treatment related effects were observed on animal body weights. Individual and mean body weights are given in Table 1.

Table 1 Individual Body Weights for all Animals with Group Means

Animal Number	Identity Number	Test GroupName	Initial BodyWeight(g)	Terminal BodyWeught(g)
5970	1	Negative control (vehicle), AOO	25.1	23.1
5958	2		24.5	24.3
5953	3		24.2	23.7
5965	4		23.4	23.7
		Mean	24.3	23.7
5973	5	100 (w/v) %  Cyclohexene in AOO	25.2	22.9
5956	6		24.4	23.9
5960	7		23.8	23.7
5967	8		23.3	22.2
		Mean	24.2	23.2
5971	9	50 (w/v) % Cyclohexene in AOO	25.6	24.3
5954	10		24.4	23.5
5959	11		24.5	24.6
5963	12		23.6	24.0
		Mean	24.5	24.1
5955	13	25 (w/v) % Cyclohexene in AOO	25.6	24.3
5957	14		24.0	24.4
5962	15		24.0	21.9
5964	16		23.6	22.6
		Mean	24.2	23.2
5966	17	Positive control, 25 (w/v) % HCA in AOO	24.9	22.8
5969	18		24.5	22.4
5972	19		24.7	23.1
5968	20		23.5	21.8
		Mean	24.4	22.5

- PROLIFERATION ASSAY

The results of the proliferation assay are summarized in Table 2. The appearance of the lymph nodes was normal in the negative (vehicle) control and test item treated groups. Larger than normal lymph nodes were observed in the positive control group.

Table2 DPM, DPN and Stimulation Index Values for all Groups

Test GroupName	Measured DPM/group	DPM	No. of Nodes	DPN	StimulationIndex Values
Background (5 (w/v) % TCA )	30.5	-	-	-	-
Negative control(AOO)	690	659.5	8	82.4	1.0
Cyclohexene100 (w/v) %	1779	1748.5	8	218.6	2.7
Cyclohexene 50 (w/v) %	854	823.5	8	102.9	1.2
Cyclohexene25 (w/v) %	980	949.5	8	118.7	1.4
Positive control(25% HCA)	7087	7056.5	8	882.1	10.7

- INTERPRATION OF OBSERVATION

There were no confounding effects of irritation or systemic toxicity at the applied concentrations and the proliferation values obtained are considered to reflect the real potential of the test item to cause lymphoproliferation in the Local Lymph Node Assay. The resulted stimulation indexes observed under these exaggerated test conditions was considered to be good evidence that Cyclohexene is a non-sensitizer.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

Under the conditions of the present assay cyclohexene, tested in a suitable vehicle, was shown to have no sensitisation potential (non-sensitizer) in the Local Lymph Node Assay.